Detection of the myristylated gag-raf transforming protein with raf-specific antipeptide sera.
Detection of the myristylated gag-raf transforming protein with raf-specific antipeptide sera.
复制标题
用 raf 特异性抗肽血清检测肉豆蔻基化的 gag-raf 转化蛋白。
DOI：
10.1016/0042-6822(85)90054-6
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发表时间：
1985
期刊：
影响因子：
3.7
通讯作者：
Oroszlan,S
中科院分区：
文献类型：
--
作者：
Schultz,AM;Copeland,TD;Mark,GE;Rapp,UR;Oroszlan,S
通过用tunicamycin抑制糖基化和用[3H]肉豆酱酸进行标记，研究了3611小鼠肉瘤病毒（MSV） gag-灌流蛋白的翻译后修饰。结果表明，p75gag - rafa是多花柱化的，而不是糖基化的，p90gag- rafa是糖基化的，而不是多花柱化的（现在称为gp90gag-raf）。gp90gag - raff的表达在转化细胞的传代过程中丢失，因此对于维持转化似乎不是必需的。从预测的v-rafsequence的不同区域合成的多肽获得了用于检测gag-raffusion蛋白的raf特异性血清。p75gag - rafaf特异性血清的免疫沉淀直接证实了推断的v- rafe序列。p759 gag-rafi在抗血清作用下呈肉鸡状并沉淀到v-rafgene的一个预测的羧基末端肽，这一事实表明，成熟蛋白代表了整个编码区。gP90gag-rafthus似乎是融合蛋白序列指定的p75gag - rafas的糖基化形式，类似于小鼠白血病病毒的pr65gag和gpr80gag。羧基末端P75gag-rafpeptide的抗血清在免疫沉淀中最有效，可用于检测c-rafgene的产物。
The post-translational modifications of thegag-raffusion proteins of the 3611 murine sarcoma virus (MSV) have been examined by inhibiting glycosylation with tunicamycin and byin vivolabeling with [3H]myristic acid. The results show that P75gag-rafis myristylated but not glycosylated and that P90gag-rafis glycosylated but not myristylated (and is now termed gp90gag-raf). gP90gag-rafexpression appeared to become lost during passage of the transformed cells, and consequently does not appear to be necessary for the maintenance of transformation.raf-specific sera for detectinggag-raffusion proteins have been obtained from synthetic peptides made from different regions of the predicted v-rafsequence. Immunoprecipitation of P75gag-rafwithraf-specific sera directly confirmed the deduced v-rafsequence. The fact that P759gag-rafis both myristylated and precipitated by antiserum to a predicted carboxyl-terminal peptide of the v-rafgene established that the mature protein represents the entire coding region. The gP90gag-rafthus appears to be a glycosylated form of P75gag-rafspecified by thegagsequences of the fusion protein, in analogy with Pr65gagand gPr80gagof murine leukemia viruses. Antiserum to the carboxyl-terminal P75gag-rafpeptide was the most efficient in immunoprecipitation, and will be useful for detecting the product of the c-rafgene.